The Core will provide proteomics and metabolomics services to the projects in the SCCOR. Methodology for proteomics analysis will be based on 2D-differential gel electrophoresis (DIGE) coupled with in-gel digestion and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI/TOF), MALDI/TOF/TOF, or liquid chromatography/tandem mass spectrometry (LC/MS/MS). Protein spots will be imaged using differential in gel electrophoresis methodology so that proteins can be quantified. All spots will be excised from the gel but identification of differentially expressed spots will be the major goal of analyses that are performed. The other spots will be stored at -70 degrees C in case other targets are discovered during subsequent research. The proteins present in gel cores will be subjected to in-gel digestion with subsequent extraction of the peptide fragments for analysis by MS. In-gel digestion will normally be conducted with trypsin. Samples will be de-salted prior to MS analyses using commercially available micro columns. The samples will then be spotted on MALDI-TOF targets or injected directly on an LC/MS system. MALDI/MS will be performed to obtain peptide maps for protein characterization. MALDI/MS/MS and LC/MS/MS will be performed for structural confirmation. The Core will also provide methodology for serum and cellular proteomics to conduct multidimensional LC/MS and LC/MS/MS of protease digests. Stable isotope dilution methodology that has been developed at the University of Pennsylvania will be used for high specificity protein quantitation. The Core will also provide diverse analytical services based primarily on liquid chromatography/tandem mass spectrometry (LC/MS/MS) methodology to quantify metabolomic biomarkers. These assays, which are based upon the use of stable isotope dilution methodology have been developed over the last several years in the Blair laboratory and are now available as routine assays to the investigators within the program. Assays that will be available include: etheno-DNA-adducts, heptanone-etheno-DNA-adducts, oxidized-DNA-adducts, glutathione adducts, chiral bioactive lipids, and protein adducts. The Core will also provide methodology for the analysis of nitric oxide, develop assays for drugs used in the SCCOR (as required), and maintain a laboratory information management system.